Site-directed mutagenesis of flavivirus E proteins
Corresponding Organization : Washington University in St. Louis
Protocol cited in 1 other protocol
Variable analysis
- Amino acid residue at position 198 of the WNV E protein (replaced with phenylalanine, and additional variants introduced using degenerate codons)
- Amino acid residue at the analogous position of the DENV1 E protein (reciprocal mutation F193T introduced)
- Amino acid residue at the analogous position of the ZIKV E protein (mutation F198T introduced)
- Not explicitly mentioned
- The structural genes (C-prM-E) of the WNV NY99 strain, DENV1 Western Pacific strain, and ZIKV strain H/PF/2013 used as templates for mutagenesis
- Use of Pfu Ultra DNA polymerase system for site-directed mutagenesis
- Transformation into Stbl2 cells and propagation at 30°C
- Sequencing of the entire C-prM-E region of each construct to ensure no additional mutations
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
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